Soils had an ability to decompose BHC when they were kept in flooded condition, and the ability was remarkably enhanced with peptone. Every tested soil was affected by the addition of peptone, and it was suggested to be due to the increase of BHC decomposers in the soil. The BHC decomposer predominating in the soil-peptone culture preferred an anaerobic condition and was heat-resistant, being able to decompose α- as well as γ-BHC. Several anaerobic bacteria were isolated from the soil-peptone culture as BHC decomposers, and these decom-posers were tentatively identified as species of Clostridium from several characteristic features.

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The volatiles from Japanese radish, processed by fermentation with rice bran, after drying (‘Hoshi Takuan’) or salting (‘Shiooshi Takuan’), have been separated and to a large extent identified by mass-spectrometric analysis. A similar study has been conducted on salted radish (‘Yamakawa Zuke’).
Isothiocyanates, constituting the major flavor characteristics of fresh radish, are rapidly catabolized. Acetic acid, alcohols, carbonyl compounds, and acetals, together with various sulfur-containing products, dominate the volatile fractions of fermented radish. A some-what different picture prevails in the case of salted radish.
The mass spectra of several sulfur compounds are discussed.

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Cell division-inducing substances in the roots of Raphanus sativus L. cv. Sakurajima were extracted with methanol and fractionated. Tobacco callus bioassay showed most of the cytokinin activity to remain in a basic fraction which is soluble to both n-butanol and hot ethyl acetate. On further successive separation, i.e., Celite column chromatography, Sephadex LH-20 column chromatography and silica gel t1c, this fraction afforded one of the components as fine needles to which most of the activity may be attributed. This was identified as trans-zeatin based on thin-layer chromatographic behavior and UV and MS spectral data.

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Aflatoxins B₁, B₂, G₁ and G₂ were quantitatively detected by high-performance liquid chromatography on a 12μ1 flow-cell in the fluorometric detector using the mobile phase of toluene system instead of chloroform, dichloromethane or methanol system. Various kinds of columns and mobile phases were tested, and fine mutual separation of all the four aflatoxins without quenching their fluorescence was achieved by using silica gel column and toluene-ethyl acetate-formic acid-methanol (89.0:7.5:2.0:1.5 v/v/v/v). The relationship between the fluorescence peak area and the amount injected was linear in the range of 0.3ng to 120ng. This method, as applied to food and feed extracts, is sensitive at the 10_??_20 ppb levels of the four kinds of aflatoxins.

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Some properties of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase in microsomes of villous and crypt cells from the jejunal and ileal epithelia of rats fed commercial pellet were studied. The optimum pH of the microsomal reductase from villi and crypts was 7.0_??_7.2 and the Km for HMG-CoA was 41.7μM. The reductase specifically required dithiothreitol for its activity. The activity was higher in ileal populations than in the jejunum. Responses of the reductase in the villous fraction to feeding cholesterol and taurocholate in combination or cholestyramine resembled those observed in crypt cells. Thus, the properties of microsomal HMG-CoA reductase in villous and crypt cells from the small intestine are similar each other, and they are possibly the same enzyme.

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Protein bodies (PB) were prepared from IR480-5-9 (10.5% protein) milled rice by destar-ching the cooked rice flour with crystalline bacterial α-amylase. Corresponding PB from raw rice prepared by treatment with analytical grade glucoamylase were more susceptible to pepsin digestion than PB from cooked rice. The undigested residue from pepsin digestion of both cooked and raw rice PB and preparations of fecal protein particles from man and rat on an IR480-5-9 cooked milled rice diet represented the core of PB. The core protein was poorer in lysine and had lower mol. wt. subunits than whole PB protein. The pepsin-digested PB preparations had higher fat content than whole PB preparation.

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The effect of several materials on viscometric behaviour of the soluble ovomucin was determined with a cone plate viscometer.
The soluble ovomucin showed a rapid increase in viscosity above 1.5mg/ml, and a high concentration of the soluble ovomucin led to gel whose viscosity was comparable to that of the insoluble ovomucin. The apparent viscosity of the soluble ovomucin decreased with an increase in NaCl concentration and upon addition of lysozyme as well as of CaCl₂. The soluble ovomucin in the presence of the sonicated β-ovomucin showed an increase in viscosity on addition of a small amount of CaCl₂.
It is assumed that a great increase in viscosity of egg white may result from removal of sodium ion and lysozyme.

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In investigations of biological active substances in metabolites of n-paraffin-utilizing microorganisms, Pseudomonas aeruginosa (Schroeter) S10B2 isolated from soil was found to produce pyoluteorin and its derivatives. One derivative was identified as 3'-nitropyoluteorin, a new metabolite of microorganisms. Some of these products were found to have anti-microbial activity in vitro and in vivo. In addition, the herbicidal activity of these products was discovered.

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A potassium chromate-tolerant bacterium was isolated from activated sludge, and the bacterium was identified as Pseudomonas ambigua G-1. The bacterium tolerated up to 2000ppm of Cr⁶⁺, 1700ppm of Cu²⁺ and 200ppm of Cd²⁺, but did not tolerate Hg²⁺. Chemical analysis indicated 86.5% uptake in the soluble fraction and 13.5% uptake in the insoluble fraction of cells. Chromate uptake distribution in the soluble fraction indicated 28.9 in microsomal fraction and 78.1% in supernatant fraction, Chromate distribution in the insoluble fraction showed 61% in lipid-fraction and 21% in polyphosphate-polysaccharide-fraction. Chromate inhibited the syntheses of protein, DNA in soluble fraction and RNA in microsomal fraction.

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Particulate alcohol dehydrogenase of acetic acid bacteria that is mainly participated in vinegar fermentation was purified to homogeneous state from Gluconobacter suboxydans IFO 12528. Solubilization of enzyme from the bacterial membrane fraction by Triton X-100 and subsequent fractionation on DEAE-Sephadex A-50 and hydroxylapatite was successful in enzyme purification. A cytochrome c-like component was tightly bound to the dehydro-genase protein and existed as an enzyme-cytochrome complex. It was also confirmed that the alcohol dehydrogenase is not a cytochrome component itself. The molecular weight of the enzyme was determined to be 150, 000, and gel electrophoresis showed the presence of three subunits having a molecular weight of 85, 000, 49, 000 and 14, 400. The smallest subunit was corresponded to the cytochrome c-like component. Ethanol was oxidized in the presence of dyes in vitro but NAD or NADP were not required as hydrogen acceptor. Unlike NAD-linked alcohol dehydrogenase in yeast or liver and other primary alcohol dehydrogenases in methanol utilizing bacteria, the enzyme from the acetic acid bacteria showed its optimum pH at fairly acidic pH.

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Crystalline 2-ketogluconate reductase in genus Acetobacter was prepared from cell free extract of Acetobacter ascendens. Crystalline enzyme was purified 13, 000-fold with a yield of 15%. Affinity chromatography on blue-dextran Sepharose 4B column successfully purified the enzyme. The enzyme was composed of three identical subunits with a molecular weight of 40, 000. Substrate specificity of 2-ketogluconate reductase from two genera of acetic acid bacteria was compared using highly purified enzyme preparations, and it was confirmed that gluconate oxidation activity of the enzyme was intrinsically weak or absent in genus Acetobacter and intense in Gluconobacter. This fact must be a useful criterion for classification of acetic acid bacteria.

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A new enzymatic method for microdetermination of ethanol has been established with particulate alcohol dehydrogenase from acetic acid bacteria and applied to the practical purposes. The enzyme had an optimum pH for ethanol oxidation at a fairly acidic region. Trace amounts of ethanol could be assayed by measuring the initial reaction rate as successful as by reading the end point of the reaction. Some advantages in using this enzyme for ethanol determination were pointed out comparing with NAD-linked alcohol dehydrogenase from yeast or horse liver. Impurity in the enzyme preparations, stability of reagents and coexistence of other substances in the assay mixture were not as critical as in NAD-linked enzyme. Acidic samples could also be directly determined for ethanol without preadjustment of sample pH.

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Investigations were carried out in order to elucidate the reason for the nutritive value reduction of browned protein by using casein labeled with U-¹⁴C-L-lysine. When the browned casein was ingested by growing rats, high radioactivity was found in a lysine derivative present in the small intestinal TCA-soluble fraction obtained 3 and 7hr after feeding. Experiments were done to identify the lysine derivative as absorption-delayed material. This material was identified by an amino acid auto-analyzer, paper chromatography and radioactive analysis as 1-deoxy-l-(ε-N-L-lysino)-D-fructose (ε-fructoselysine), which accounted for about 70% of the total radioactivity in the small intestinal TCA-soluble fraction obtained 7hr after feeding. Lysine which was found after 3hr, was disappeared from the intestinal TCA-soluble fraction taken 7hr after feeding, but ε-fructoselysine remainded. Its content as acetate found 7hr after one-dosal feeding of 600mg browned labeled casein (700, 000 dpm) was 18.3 and 19.0mg/each rat, respectively. These values accounted for about 40% of the lysine content in the ingested browned casein.

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Kinetic study of glycolaldehyde dehydrogenase was performed with isozymes of Escherichia coli B. The reaction equilibrium of isozyme A was estimated to lie in glycolate formation, while those of isozymes B and C were in glycolaldehyde formation. Isozyme A was released from cells with osmotic shock, while the others were not. Isozymes B and C were found in cytoplasmic fraction. Some reversal mutants derived from WG3 strain (one of vitamin B₆ auxotrophs) acquired ability to produce isozyme C. Based on these results, the non-involve-ment of isozyme A in vitamin B₆ biosynthesis was discussed.

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1. Quantitative changes in fractional catabolic and synthetic rates of the myosin-actin pool in rat muscle under starvation and refeeding, during growth or after treatment with hydrocortisone were studied by estimating urinary excretion of N_??_-methylhistidine (3-methyl-bistidine; Me-His).
2. Following deprivation of food, urinary Me-His output increased from 0.35mg/day to 0.45mg/day during first 2 day in spite of decreasing body Me-His pool. This high rate of Me-His excretion was maintained for the following 4 days of starvation and then decreased. When rats were refed a 20% casein diet after 10 days of starvation, Me-His excretion con-tinued to decrease even after 3 days of refeeding. On the fifth day of refeeding, it began to rise progressively. During starvation, fractional catabolic rate of myosin-actin was about 3.7%/day in comparison with 2.6%/day of fed rats. After refeeding, the fractional catabolic rate decreased rapidly to a minimum value of 1.7%/day on the third day. After that, it reached to a value of 2.6%/day of fed rats. On the other hand, fractional synthetic rate of myosin-actin dropped immediately after fasting and the low rate of about 0.4%/day was maintained during starvation period. Fractional synthetic rate recovered quickly after refeeding.
3. Urinary output of nitrogen and creatinine rose quickly on the first day after ad-ministration of hydrocortisone and on the second day it fell to their normal value. While Me-His excretion increased after injection of hydrocortisone up to 0.52mg/day on the second day and this high excretion rate remained until the following day. From these results, it was shown that administration of hydrocortisone to rats enhances catabolism and reduces synthesis of myosin-actin. The results also show that the effect of this hormone on myofibrillar protein catabolism appears to last longer than its effect on nitrogen metabolism in the whole body judged from urinary nitrogen output.
4. Fractional rates of catabolism and synthesis of rat myosin-actin were 3.3%/day (half-life of 21 days) and 7.2%/day, respectively, at the growth stage of 129g body weight. These rates were 2.3%/day (half-life of 30 days) and 2.8%/day, respectively, at the mature stage of 363g body weight.
5. Under the dietary conditions in this experiment, fractional synthetic rate changed far more dramatically than catabolic rate. This suggests that mass of muscle protein is primarily regulated by the rate of synthesis, although the rate of catabolism should not be neglected.

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Localization of α-galactosidase in an alkalophilic strain of Micrococcus was investigated in relation to the cell membrane as a permeability barrier. The most α-galactosidase ap-pered to be intracellular; only about 4% of α-galactosidase was released by lysozyme or freeze-thaw treatments of the whole cells. The enzyme activity was not inhibited by treatment of the whole cells with diazo-7-amino-1, 3-naphthalene disulfonic acid (NDS) which penetrated the cell wall but not the cytoplasmic membrane. The enzyme activity of the whole cells increased about four-fold by toluene-acetone treatment which caused an alteration in the membrane permeability. The enzyme in such cells became to be relatively sensitive to pH. These results showed that cell membrane played a protective role as a permeability barrier against alkaline environment.

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Quinolinate phosphoribosyltransferase (EC 2.4.2.19) was purified and crystallized from cell-free extracts of Alcaligenes eutrophus nov. subsp. quinolinicus, IAM 12305 which was isolated in our laboratory from soil. The enzyme was labile in the cold, and all purification steps were performed at room temperature (10_??_15°C). The crystalline enzyme was certified to be homogeneous by ultracentrifugal analysis and starch-gel electrophoresis. On poly-acrylamide gel disc electrophoresis, the crystalline enzyme showed a multiple profile, but it showed a single band by addition of a certain amount of glycerol and 2-mercaptoethanol. The adding effect of these compounds was discussed.

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Native glycinin was split by trypsin in limited regions under high ionic strength con-dition and converted into glycinin-T. Initially, the intermediary subunits of glycinin con-verted into fragments of 48, 000 (IST-1) and 44, 500 (IST-2) molecular weights. These frag-ments were transitory intermediates and then they were cleaved yielding fragments of 32, 500 (IST-3) and 29, 000 (IST-4) molecular weights. Analyses of the mole fractions of the frag-ments revealed that IST-2 was rapidly split into IST-4 followed by splitting of IST-1 into IST-3.
Subsequently, glycinin-T was dissociated with 6_M urea and subjected to gel filtration and ion exchange chromatography in order to isolate IST-3 and IST-4. Acidic-urea gel electrophoresis of the isolated IST showed that IST-3 contained B₁, B₂ and B₃ subunits, whereas IST-4 contained B₄ subunit. These results indicated that IST-3 was formed from IS-I and IS-2, and IST-4 from IS-3.

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To investigate the constituents of Z-disk, reconstitution of Z-disk by incubating some proteins released from myofibrils by CAF (Ca²⁺-activated factor) with Z-disk-extracted fiber bundles was carried out and examined with electron microscope. The materials released from myofibrils by CAF have been bound in Z-disk region, and Z-disk extracted from myo-fibrils with a low ionic strength solution has been reconstituted. On the other hand, Z-disk removed from myofibrils by CAF has not been reconstituted by the same way.

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To investigate the exact protein constituents of Z-disk on the basis of the success or fail-ure of reconstitution of Z-disk, proteins released from myofibrils by CAF(Ca²⁺-activated factor) were fractionated, the Z-disk was reconstituted by incubating individual fractions with Z-disk-extracted fiber bundles and the proteins in each fraction were analyzed by polyacrylamide gel electrophoresis.
Released materials from myofibrils by CAF were divided into three fractions, A, B and C, in the order of elution from a Sepharose 6B column. The materials in Fractions A and B have been bound in the Z-disk region, and the Z-disk extracted from myofibrils in a low ionic strength solution has been reconstituted. The Z-disk reconstituted by incubating the materials in Fraction A with Z-disk-extracted myofibrils seems to have a structure similar to the intact Z-disk. Fraction A consisted principally of proteins having subunit molecular weights near 100, 000 and 34, 000 daltons.

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The volatile basic fraction concentrates were prepared from heated, and raw shoyu, separately. Each concentrate was successively analyzed by combined gas chromatography-mass spectrometry. In the above two concentrates, 35 components were identified, 27 of which have not been reported previously for their presence in shoyu. The identified compounds were 24 pyrazines, 5 pyridines, 2 oxazoles and 4 other compounds. Furthermore, the basic compounds of both heated and raw shoyu were quantitatively analyzed by GC, and the concentrations and odor units of major pyrazines which contained simple alkyl groups as the side chain were also determined. Consequently, the quantity of the basic compounds was found to increase during pasteurization; especially the total quantity of the above major pyrazines increased remarkably, Therefore, it is concluded that these confirmed pyrazines which have very low odor threshold values play an important role in the so-called “heated” flavor which is brought about by pasteurization of raw shoyu in the course of production. In addition, organoleptically, the basic compounds are considered to be indispensable to the shoyu flavor.

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SY-1 (I) is an antibiotic produced by Streptoinyces albus ATCC 21838 as a concurrent minor component of salinomycin. The antibiotic was isolated in the form of sodium salt, C₄₂H₆₉O₁₀Na. The structure of I has been established by the NMR and mass spectral evidences as well as by direct comparison of the 18, 19-dihydroSY-1 methyl ester (IV) with 18, 19-dihydro-20-deoxysalinomycin methyl ester (VII). I showed a weak inhibitory activity against gram-positive bacteria and was also effective in the treatment of coccidial infection in chickens.

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SY-1 (20-deoxysalinomycin), a monocarboxylic polyether antibiotic closely related to salinomycin, caused a rapid release of previously accumulated alkali metal cations by valino-mycin or monazomycin in rat liver mitochondria, and simultaneously reversed swelling of mitochondria.
With a strict specificity for substrate and cation, SY-1 exhibited a property of inhibiting mitochondrial functions such as substrate oxidation, oxidative phosphorylation and ATP hydrolysis induced by valinomycin or monazomycin. In comparative study with salinomycin, SY-1 was found to be more effective on the mitochondrial functions than salinomycin.
On the basis of the results so far obtained, the inhibitory effect of SY-1 on mitochondria is interpreted as a result of interaction with essential cations, especially with. K⁺, in mitochondria.

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A new method for the selective determination of inorganic arsenic (III), (V) and organic arsenic has been performed. Each chemical form of arsenic in biological materials was solubilized in 6N hydrochloric acid. Inorganic arsenic (III) was extracted into toluene from 9N hydrochloric acid solution, and inorganic arsenic (V) was also extracted after reduction by potassium iodide. Organic arsenic was retained in hydrochloric acid solution. After arsenic in toluene solution was back-extracted into water, aqueous or hydrochloric acid solution containing arsenic was wet digested with a mixture of HNO₃, H₂SO₄ and HClO₄, and arsenic was determined by atomic absorption spectrophotometry. The proposed method was applied to some kinds of algaes, shark muscle and orchard leaves (NBS SRM 1571). 90_??_100% of arsenic in algaes (Wakame, Arame, Kazime) and shark muscle was organic, while all the arsenic in orchard leaves was inorganic.

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Escherichia coli cells acquired supersensitivity to various β-lactam antibiotics by dacA mutation, a defect in D-alanine carboxypeptidase IA activity. The mutant cells were rather less sensitive to mecillinam than the dacA⁺ cells. This mutation did not result in either thermo-sensitivity of cell growth or appreciable increase of the generation times in usual rich media, but the resulting appearance of supersensitivity to β-lactam antibiotics suggests that the cell wall or envelope of this mutant is somewhat abnormal and thus that D-alanine carboxypeptidase IA is involved in cell wall or envelope synthesis.

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